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Creation of a Stable P19 Cell Line Producing Pts2-Egfp



Mojbafan M1 ; Ghaedi K2 ; Razavi S3 ; Karamali F4 ; Karbalaii K4 ; Tanhaie S4 ; Rabiee F4 ; Esfahani MHN5 ; Baharvand H6
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Authors Affiliations
  1. 1. Department of Biology, School of Science, University of Isfahan, Isfahan, Iran
  2. 2. School of Science, University of Isfahan, Department of Cell and Molecular Biology, Isfahan, Iran
  3. 3. Department of Anatomy, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
  4. 4. Department of Cell and Molecular Biology, Royan Institute for Animal Biotechnology, Iranian Academic Center for Education, Culture and Research (ACECR), Tehran, Iran
  5. 5. Department of Developmental Biology, Royan Institute for Stem Cell Biology and Technology, Iranian Academic Center for Education, Culture and Research (ACECR), Isfahan, Iran
  6. 6. Department of Stem Cells and Developmental Biology, Royan Institute for Stem Cell Biology and Technology, Iranian Academic Center for Education, Culture and Research (ACECR), Tehran, Iran

Source: Journal of Isfahan Medical School Published:2010

Abstract

Background: P19 cells are mouse embryonic carcinoma cells which contain pluripotent ability, like stem cells, to differentiate into different cell lines. There are several properties for this cell line that make it a valuable cell model for study of developmental stages. Methods: At the first step, PTS2-EGFP coding sequence which was cloned in pUcD2.hygro vector was used for transfection in to P19 cells. As the plasmid contained hygromycin (Hygror) resistance gene, stable cells were selected using hygromycin as an antibiotic. Stable transformed cells were characterized by RT-PCR and immunostaining analyses. Findings: RT-PCR results indicated EGFP was expressed in these cells. Moreover immunocytochemical analysis of the cells confirmed the preserved pluripotency states of transfected cells. Conclusion: As P19 Cells are able to differentiate into several cell lines, using this stable cell line we will able to chase the molecular kinetics of peroxisomes.
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